Yeh Jeng-Hsien, Chou Chiang-Ting, Chen I-Shu, Lu Ti, Lin Ko-Long, Yu Chia-Cheng, Liang Wei-Zhe, Chang Hong-Tai, Kuo Chun-Chi, Ho Chin-Man, Chang Wen-Teng, Shieh Pochuen, Jan Chung-Ren
Pathology and Laboratory Medicine Department, Kaohsiung Veterans General Hospital, Kaohsiung 81362, Taiwan, Republic of China.
Department of Biological Science and Technology, Chung Hwa College of Medical Technology, Tainan 71703, Taiwan, Republic of China.
Chin J Physiol. 2017 Feb 28;60(1):32-40. doi: 10.4077/CJP.2017.BAF447.
Thymol is a phenolic compound that affects physiology in different cell models. However, whether thymol affects Ca²⁺ homeostasis in prostate cancer cells is unknown. The action of this compound on cytosolic Ca²⁺ concentrations ([Ca²⁺]i) and viability in PC3 human prostate cancer cells was explored. The results show that thymol at concentrations of 100-1500 μM caused [Ca²⁺]i rises in a concentration-dependent manner. Removal of extracellular Ca²⁺ reduced thymol’s effect by approximately 80%. Thymol-induced Ca²⁺ entry was confirmed by Mn²⁺ entry-induced quench of fura-2 fluorescence, and was inhibited by approximately 30% by Ca²⁺ entry modulators (nifedipine, econazole, SKF96365), and the protein kinase C (PKC) inhibitor GF109203X. In Ca²⁺-free medium, treatment with the endoplasmic reticulum Ca²⁺ pump inhibitor thapsigargin abolished thymol-induced [Ca²⁺]i rises. Treatment with thymol also abolished thapsigargin-induced [Ca²⁺]i rises. Thymol-induced Ca²⁺ release from the endoplasmic reticulum was abolished by the phospholipase C (PLC) inhibitor U73122. Thymol at 100-900 μM decreased cell viability, which was not reversed by pretreatment with the Ca²⁺ chelator 1,2-bis(2-aminophenoxy) ethane-N,N,N’,N’-tetraacetic acid-acetoxymethyl ester (BAPTA/AM). Together, in PC3 cells, thymol induced [Ca²⁺]i rises by inducing PLC-dependent Ca²⁺ release from the endoplasmic reticulum and Ca²⁺ entry via PKC-sensitive store-operated Ca²⁺ channels and other unknown channels. Thymol also induced Ca²⁺-dissociated cell death.
百里香酚是一种酚类化合物，在不同细胞模型中会影响生理功能。然而，百里香酚是否会影响前列腺癌细胞中的钙离子稳态尚不清楚。本研究探讨了该化合物对PC3人前列腺癌细胞胞质钙离子浓度（[Ca²⁺]i）及细胞活力的作用。结果表明，浓度为100 - 1500 μM的百里香酚可引起[Ca²⁺]i浓度依赖性升高。去除细胞外钙离子可使百里香酚的作用降低约80%。通过锰离子进入诱导的fura - 2荧光淬灭证实了百里香酚诱导的钙离子内流，钙离子内流调节剂（硝苯地平、益康唑、SKF96365）和蛋白激酶C（PKC）抑制剂GF109203X可使其抑制约30%。在无钙离子培养基中，用内质网钙离子泵抑制剂毒胡萝卜素处理可消除百里香酚诱导的[Ca²⁺]i升高。用百里香酚处理也可消除毒胡萝卜素诱导的[Ca²⁺]i升高。磷脂酶C（PLC）抑制剂U73122可消除百里香酚诱导的内质网钙离子释放。100 - 900 μM的百里香酚可降低细胞活力，钙离子螯合剂1,2 - 双（2 - 氨基苯氧基）乙烷 - N,N,N',N' - 四乙酸 - 乙酰氧甲酯（BAPTA/AM）预处理不能逆转这种作用。总之，在PC3细胞中，百里香酚通过诱导PLC依赖的内质网钙离子释放以及通过PKC敏感的储存 - 操纵性钙离子通道和其他未知通道引起钙离子内流，从而导致[Ca²⁺]i升高。百里香酚还可诱导钙离子解离的细胞死亡。
